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Venous blood was drawn for determination of blood glucose, HbA1c and serum concentrations of total cholesterol, HDL-C and triglycerides at baseline, and then at least twice a year. Plasma glucose was measured by the glucokinase method, and HbA1c by ion-exchange high-performance liquid chromatography. The Japan Diabetes Society (JDS) has standardized several HbA1c assays with the international standard value adjusted by the equation of HbA1c (JDS) (%) plus 0.4%. Serum insulin was measured by an enzyme immunoassay, and total cholesterol, triglycerides, HDL-C, white blood cells, red blood cells, hematocrit (Ht), blood urea nitrogen (BUN), serum creatinine, uric acid, total protein and albumin by established methods. Blood pressure was measured with a mercury sphygmomanometer using a cuff of appropriate size. http://www.selleckchem.com/products/gsk1120212-jtp-74057.html Diastolic blood pressure was determined as Korotkoff phase?V. Body mass index was calculated as weight in kilograms?/?(height in meters)2. Microangiopathy (retinopathy, nephropathy and neuropathy), macroangiopathy (ischemic http://www.selleckchem.com/products/MDV3100.html heart disease [IHD]), stroke and peripheral vascular disease [PVD]) were assessed at baseline, and then annually. Funduscopic examinations were carried out on dilated pupils by experienced ophthalmologists using direct ophthalmoscopy. Retinopathy status was assessed by the Japanese Diabetes Complication Study method and classified into five stages: stage 0: no retinopathy; stage 1: dot hemorrhages, hemorrhages or hard exudates; stage 2: soft exudates; stage 3: IRMA or venous deformities; stage 4: neovascularization, preretinal proliferative tissues, vitreous hemorrhages or retinal detachment. Diabetic maculopathy was assessed according to findings of hemorrhages, local edema, hard exudates and diffuse edema at macular areas. Uncorrected and corrected visual acuities, the occurrence of cataract, corneal opacity, glaucoma, age-related macular degeneration, http://www.selleck.cn/products/pd-1-pd-l1-inhibitor-2.html laser photocoagulation, cataract operations and vitrectomy were assessed. Urinary albumin was measured by immunological assay. Mean urinary albumin-to-creatinine ratio (ACR; ?g/mg creatinine) in two or three successive urinalyses was used to classify diabetic nephropathy as no nephropathy (ACR?